PMID: 9168145Apr 26, 1997Paper

Comparison of ouabain-sensitive and -insensitive Na/K pumps in HEK293 cells

Biochimica Et Biophysica Acta
J KockskämperH G Glitsch

Abstract

The Na/K pump current I(p) of single HEK293 cells either untransfected (endogenous I(p)) or transfected with the alpha1 subunit of the rat Na/K pump (exogenous I(p)) was investigated in Na-containing solution by means of whole-cell recording at 30 degrees C. The endogenous I(p) was irreversibly blocked by 10(-4) M ouabain or 2 x 10(-4) M dihydro-ouabain (DHO). Its density amounted to 0.33 pA pF(-1) at 0 mV and 5.4 mM K(o). It was half maximally activated at 1.5 mM K(o) and increased linearly with depolarization over the entire voltage range studied (-80 to +60 mV). In contrast, HEK293 cells stably transfected with cDNA for the cardiac glycoside-resistant alpha1 subunit of the rat Na/K pump showed an I(p) in the presence of 10(-4) M ouabain and 2 x 10(-4) M DHO, respectively. This exogenous I(p) was reversibly blocked by 10(-2) M ouabain. Half maximal activation of the exogenous I(p) occurred at 1.7 mM K(o). Its amplitude increased linearly with depolarization at negative voltages but remained almost constant at positive membrane potentials. Comparison with the I(p) of isolated rat cardiac ventricular myocytes strongly suggests that the exogenous I(p) in HEK293 cells is generated by the alpha1 subunit of the rat Na/K pump since ...Continue Reading

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Citations

Mar 30, 2001·The Journal of General Physiology·P De WeerR F Rakowski
Mar 17, 2004·The Journal of General Physiology·Pablo Artigas, David C Gadsby
Sep 16, 2015·Proceedings of the National Academy of Sciences of the United States of America·Beata UjvariThomas Madsen
Jul 24, 1998·Biochimica Et Biophysica Acta·U Eckstein-LudwigW Schwarz
May 29, 2019·PloS One·Michael SolarskiGerold Schmitt-Ulms
Nov 18, 2016·Proceedings. Biological Sciences·Shabnam MohammadiAlan H Savitzky

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